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Catalog # HK323

MBL, Human, ELISA kit

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Mannose Binding Lectin (MBL) is a C-type lectin that is an important element in innate immunity. MBL belongs to the collectin family of proteins that consists of a collagen-like domain and a carbohydrate recognition domain (CRD). Through this, MBL recognizes carbohydrates (e.g. mannose and N-acetylglucosamine) on pathogens. MBL forms several sizes of oligomers that are composed of subunits of three identical 32 kDa polypeptides. MBL is a pattern recognition receptor, and upon recognition of the infectious agent, MBL triggers the activation of the lectin-complement pathway through attached C1r/C1s-like serine proteases, designated MASP. The MBL-MASP complex proteolytically cleaves C4, C2 and C3. The lectin pathway is antibody and C1q-independent and is, therefore, independent of the classical and alternative complement activation pathway. MBL is synthesized by hepatocytes and has been isolated from the liver or serum of several vertebrate species. Normal human plasma contains MBL concentrations ranging from 10 to 5,000 ng/ml. Up to 12% of healthy Caucasian blood donors have MBL concentrations below 100 ng/ml. Low plasma concentrations have been associated with an inherited defect in opsonization. The MBL concentration is enhanced in infectious diseases. Measuring of MBL is indicated in recurrent infections (especially in children), primary/secondary immunodeficiencies, artherosclerosis/coronary heart disease, cystic fibrosis, transplantation, autoimmune diseases (SLE/Rheumatoid arthritis) and habitual abortion. MBL measurement is not affected by the presence of antibodies against mannan. In the human MBL assay, any influence of the classical pathway has been eliminated by using a special MBL-binding buffer, which inhibits the binding of C1q to immunocomplexes and disrupts the C1 complex, while leaving the function of the MBL complex intact.

Application
The human MBL (Lectin assay) ELISA kit is to be used for the in vitro quantitative determination of human MBL in serum, plasma and cell culture supernatant samples.

Principle
The human MBL ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 4 hours. The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. After activation, samples and standards are captured by solid bound mannan. Biotinylated tracer antibody will bind to captured MBL. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the MBL standards (log). The MBL concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Product type
Assays
Amount
1 x 96 det., 2 x 96 det.
Standard range
0.41-100 ng/ml
Detection level
0.41 ng/ml
Working volume
100 µl/well
Species
Human
Alias
Mannose- or mannan-binding protein (MBP)
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Disease
Infectious diseases
Safety Data Sheet Assay Manual
HK323
405 kb

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